Background: Pseudomonas aeruginosarepresents a major concern in nosocomial infections.  It possesses manyvirulence factors that aid its pathogenicity and antimicrobial resistance. Non-antibiotic therapy against this pathogen includes probiotics, phages, phytomedicine and other new modalities. These agents alone or in combination with antibiotics can be highly effective against these virulent strains. Objectives: To investigate the in-vitro effect exerted by the probiotic “Lactobacillus plantarum" on two virulence factors of Pseudomonas aeruginosa (pyocyanin pigment production and elastase enzyme activity). Methods: The study was carried out on 40 Pseudomonas aeruginosa strains isolated from pus collected from wound infection using sterile swabs. The effect of Lactobacillus plantarum on pyocyanin production and the elastolytic activity of Pseudomonas aeruginosa were detected using pyocyanin extraction method and Human Elastase ELISA Kit respectively before and after the addition of Lactobacillus plantarum. Results: Among the forty strains investigated, the pyocyanin production and elastolytic activity of Pseudomonas aeruginosa isolates were significantly decreased after addition of Lactobacillus plantarum. Conclusions: Lactobacillus plantarum has an important invitro anti-pathogenic effect against Pseudomonas aeruginosa as it significantly interferes with two of its main virulence factors (pyocyanin & elastase).